Oropesa-Ávila M, Andrade-Talavera Y, Garrido-Maraver J, Cordero M D, de la Mata M, Cotán D, Paz M V, Pavón A D, Alcocer-Gómez E, de Lavera I, Lema R, Zaderenko A P, Rodríguez-Moreno A, Sánchez-Alcázar J A
Centro Andaluz de Biología del Desarrollo (CABD-CSIC-Universidad Pablo de Olavide), and Centro de Investigación Biomédica en Red Enfermedades Raras, Instituto de Salud Carlos III, Sevilla, Spain.
Laboratorio de Neurociencia Celular y Plasticidad, Universidad Pablo de Olavide, Sevilla, Spain.
Cell Death Dis. 2014 Aug 14;5(8):e1369. doi: 10.1038/cddis.2014.332.
Apoptosis is characterized by degradation of cell components but plasma membrane remains intact. Apoptotic microtubule network (AMN) is organized during apoptosis forming a cortical structure beneath plasma membrane that maintains plasma membrane integrity. Apoptotic cells are also characterized by high reactive oxygen species (ROS) production that can be potentially harmful for the cell. The aim of this study was to develop a method that allows stabilizing apoptotic cells for diagnostic and therapeutic applications. By using a cocktail composed of taxol (a microtubule stabilizer), Zn(2+) (a caspase inhibitor) and coenzyme Q10 (a lipid antioxidant), we were able to stabilize H460 apoptotic cells in cell cultures for at least 72 h, preventing secondary necrosis. Stabilized apoptotic cells maintain many apoptotic cell characteristics such as the presence of apoptotic microtubules, plasma membrane integrity, low intracellular calcium levels and mitochondrial polarization. Apoptotic cell stabilization may open new avenues in apoptosis detection and therapy.
细胞凋亡的特征是细胞成分降解,但质膜保持完整。凋亡微管网络(AMN)在细胞凋亡过程中形成,在质膜下方形成一种皮质结构,维持质膜完整性。凋亡细胞的另一个特征是产生高活性氧(ROS),这可能对细胞有潜在危害。本研究的目的是开发一种方法,用于稳定凋亡细胞以用于诊断和治疗应用。通过使用由紫杉醇(一种微管稳定剂)、Zn(2+)(一种半胱天冬酶抑制剂)和辅酶Q10(一种脂质抗氧化剂)组成的混合物,我们能够在细胞培养物中稳定H460凋亡细胞至少72小时,防止继发性坏死。稳定的凋亡细胞保持许多凋亡细胞特征,如凋亡微管的存在、质膜完整性、低细胞内钙水平和线粒体极化。凋亡细胞的稳定可能为细胞凋亡检测和治疗开辟新途径。